| 质粒名称: | LentiRCas9-CUG |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 104183 |
| 存储实验室: | Eugene Yeo |
|
载体骨架基本信息 |
|
| 载体名称: | LentiCRISPR v2 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 慢病毒，CRISPR |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | U6-CUGsgRNA, EFS-PIN-dCas9 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | LKO1.5 |
| 3' 测序引物 | T3 |
|
文献引用方法 |
|
| 材料和方法部分: | LentiRCas9-CUG was a gift from Eugene Yeo (Addgene plasmid # 104183 ; http://n2t.net/addgene:104183 ; RRID:Addgene_104183) |
| 参考文献部分: | Elimination of Toxic Microsatellite Repeat Expansion RNA by RNA-Targeting Cas9. Batra R, Nelles DA, Pirie E, Blue SM, Marina RJ, Wang H, Chaim IA, Thomas JD, Zhang N, Nguyen V, Aigner S, Markmiller S, Xia G, Corbett KD, Swanson MS, Yeo GW. Cell. 2017 Aug 24;170(5):899-912.e10. doi: 10.1016/j.cell.2017.07.010. Epub 2017 Aug 10. 10.1016/j.cell.2017.07.010 PubMed 28803727 |
买家导航
LentiRCas9-CUG载体 LentiRCas9CUG质粒基本信息
LentiRCas9-CUG质粒图谱 LentiRCas9CUG载体图谱和LentiRCas9-CUG载体序列 LentiRCas9CUG质粒序列多克隆位点信息
LentiRCas9-CUG质粒 LentiRCas9CUG载体简介
LentiRCas9-CUG质粒序列 LentiRCas9CUG载体序列
LOCUS LentiRCas9-CUG 13752 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Lentiviral transfer vector that carries U6-driven sgRNA targeting
CUG repeats using a modified scaffold (Chen et al. Cell 2013) and
CMV-driven PIN-dCas9. Derived from LentiCRISPR v2 (Zhang lab).
ACCESSION .
VERSION .
KEYWORDS LentiRCas9-CUG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13752)
AUTHORS Batra R, Nelles DA, Pirie E, Blue SM, Marina RJ, Wang H, Chaim IA,
Thomas JD, Zhang N, Nguyen V, Aigner S, Markmiller S, Xia G, Corbett
KD, Swanson MS, Yeo GW
TITLE Elimination of Toxic Microsatellite Repeat Expansion RNA by
RNA-Targeting Cas9.
JOURNAL Cell. 2017 Aug 24;170(5):899-912.e10. doi:
10.1016/j.cell.2017.07.010. Epub 2017 Aug 10.
PUBMED 28803727
REFERENCE 2 (bases 1 to 13752)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..13752
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 382..580
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 598..778
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 825..950
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1443..1676
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2203..2320
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2371..2611
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 2371..2391
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2542..2561
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(2748..2764)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2748..2764)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(2801..2821)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2801..2819)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2881..3092
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 3720..7820
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 7824..7850
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 7869..7889
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7896..7916
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7938..7994
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 7995..8591
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 8488..8508
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 8607..9195
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8660..8680)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(9078..9089)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 9267..9500
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(9526..9543)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 9532..9756
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 9802..10230
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9889..9908)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 10099..10120
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(10239..10259)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 10244..10573
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 10424..10559
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 10486..10505
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 10621..10668
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 10687..11061
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 11191..11312
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(11228..11247)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 11282..11301
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(11361..11377)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11361..11377)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(11374..11396)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 11385..11401
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(11409..11439)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11454..11475
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(11592..11609)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(11763..12351)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(11843..11862)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(12522..13382)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 13145..13164
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(13383..13487)
/gene="bla"
/label=AmpR promoter
primer_bind complement(13562..13581)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
2341 cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat
2401 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
2461 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
2521 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
2581 ttcttggctt tatatatctt gtggaaagga cgaaacaccg agcagcagca gcagcagcag
2641 gtttaagagc tatgctggaa acagcatagc aagtttaaat aaggctagtc cgttatcaac
2701 ttgaaaaagt ggcaccgagt cggtgctttt tttcctgcag cccgggggat ccactagttc
2761 tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg
2821 cgcgcgaatt cgctagctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt
2881 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgat
2941 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
3001 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
3061 tttttcgcaa cgggtttgcc gccagaacac aggaccggtt ctagagcgct ctcgagagaa
3121 ccatgcagat ggagctcgaa atcaggccgc tgttcctcgt gccggacact aatggtttta
3181 tagatcactt ggcgtccttg gctagacttc tggaaagccg aaagtatata ttggtagtgc
3241 cgttgattgt aattaacgaa ttggatgggt tggcgaaagg acaagagact gatcacagag
3301 caggaggcta cgcgagggtc gtccaagaga aggcgcgaaa aagcatcgag ttcctggagc
3361 agcgatttga gagcagggac tcatgcctga gagccctcac gtcacggggg aacgagctgg
3421 agtccatcgc tttccgaagt gaagacatta cgggccaact tgggaataat gatgacctca
3481 tcttgtcctg ctgcctgcac tactgcaagg acaaggctaa ggacttcatg cctgcctcca
3541 aggaggagcc tatccgattg ttgagggaag tagtactttt gacggacgac cgcaacctcc
3601 gggtaaaggc gctgactcga aatgtcccag taagggatat accggcgttc cttacatggg
3661 ctcaagtagg gagtggaagt gagacaccgg gaacctcaga gagcgccacg ccagaaagcg
3721 acaagaagta cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca
3781 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
3841 gcatcaagaa gaacctgatc ggcgccctgc tgttcgacag cggagaaaca gccgaggcca
3901 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
3961 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
4021 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
4081 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
4141 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
4201 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
4261 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
4321 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
4381 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga
4441 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
4501 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
4561 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
4621 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
4681 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
4741 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
4801 acatcgatgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
4861 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
4921 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
4981 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
5041 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
5101 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
5161 tggacaaggg cgccagcgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
5221 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtaca
5281 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
5341 gcgagcagaa aaaagccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
5401 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
5461 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
5521 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
5581 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
5641 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
5701 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
5761 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
5821 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
5881 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
5941 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
6001 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
6061 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
6121 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
6181 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gacgctatcg
6241 tgcctcagag ctttctgaag gacgactcca tcgataacaa agtgctgact cggagcgaca
6301 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
6361 actggcgcca gctgctgaat gccaagctga ttacccagag gaagttcgac aatctgacca
6421 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
6481 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
6541 agtacgacga gaacgacaaa ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
6601 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
6661 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
6721 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
6781 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
6841 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
6901 tgatcgagac aaacggcgaa acaggcgaga tcgtgtggga taagggccgg gactttgcca
6961 ccgtgcggaa agtgctgtct atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
7021 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgacaag ctgatcgcca
7081 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
7141 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
7201 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
7261 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
7321 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
7381 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
7441 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
7501 acaaacacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
7561 tggccgacgc taatctggac aaggtgctga gcgcctacaa caagcacaga gacaagccta
7621 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
7681 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
7741 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
7801 tgtctcagct gggaggcgac gcctatccct atgacgtgcc cgattatgcc agcctgggca
7861 gcggctcccc caagaaaaaa cgcaaggtgg aagatcctaa gaaaaagcgg aaagtggacg
7921 tgttcgaagg gtccggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag
7981 agaatcctgg accgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
8041 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
8101 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
8161 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
8221 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
8281 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
8341 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca
8401 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
8461 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
8521 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
8581 ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
8641 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
8701 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
8761 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
8821 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
8881 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
8941 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
9001 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
9061 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
9121 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
9181 gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag
9241 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
9301 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
9361 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
9421 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
9481 agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt
9541 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
9601 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
9661 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
9721 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
9781 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
9841 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
9901 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
9961 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
10021 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
10081 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
10141 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
10201 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
10261 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
10321 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
10381 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
10441 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
10501 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
10561 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
10621 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
10681 taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
10741 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
10801 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
10861 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
10921 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
10981 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
11041 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
11101 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
11161 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
11221 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
11281 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
11341 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
11401 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
11461 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
11521 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
11581 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
11641 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
11701 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
11761 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
11821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
11881 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
11941 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
12001 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
12061 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
12121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
12181 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
12241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
12301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
12361 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
12421 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
12481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
12541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
12601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
12661 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
12721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
12781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
12841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
12901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
12961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
13021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
13081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
13141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
13201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
13261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
13321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
13381 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
13441 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
13501 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
13561 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
13621 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
13681 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
13741 gatatacgcg tt
//
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